We found that intact speech evoked larger negativity between 280–527-ms than vocoded speech, suggestive of more robust semantic prediction for the intact signal. For visual reliability, we found that greater cross-modal ERP suppression occurred for clear than blurred videos prior to sound onset and for the P2 ERP. Additionally, the later semantic-related negativity tended to be larger for clear than blurred videos. These results suggest that the cross-modal effect is largely confined to suppression of early auditory networks with weak effect on networks associated with semantic prediction. However, the semantic-related visual effect on the late negativity may have been tempered by the vocoded signal’s high-reliability.The efficacy of audiovisual (AV) integration is reflected in the degree of cross-modal suppression of the auditory event-related potentials , while stronger semantic encoding is reflected in enhanced late ERP negativities . We hypothesized that increasing visual stimulus reliability should lead to more robust AV-integration and enhanced semantic prediction, reflected in suppression of auditory ERPs and enhanced N450, respectively. EEG was acquired while individuals watched and listened to  Looking at a talker’s mouth during conversations can boost comprehension of the speech signal, especially in noisy situations ,2. Past A consistent finding in audiovisual (AV) studies, is the cross-modal suppressive effect on AEPs ,11,12,13vocoded nonwords (see Methods for explanation about vocoding), Shatzer et al.  with no known hearing problems or language deficits. They were recruited from the UC Davis campus or local community and tested in the UC Davis Center for Mind and Brain. Participants received monetary remuneration in exchange for completing the study. Informed consent was obtained in accordance with protocols reviewed and approved by the UC Davis Institutional Review Board (IRB) on 26 March 2020 (IRB #725055-1). All procedures followed the approved guidelines of the IRB.Twenty-seven participants completed the study. Two participants were excluded from the final analysis because they did not satisfy specific criteria assuring task vigilance: one person acknowledged that they avoided looking at the blurred videos, while another did not respond to 45% of the probe trials (see below). Of the 25 remaining participants, demographic information for two subjects was lost/destroyed and thus not available. Demographic data are thus based on 23 participants . Despite the loss of demographic information on two participants, behavioral and EEG data are available for all 25. Participants (www.videostudiopro.com), such that each clip had about 10 silent frames before and after the speech utterance. Audio and silent video stimuli were extracted and saved separately for subsequent manipulation.Speech materials included forty monosyllabic animate  and forty monosyllabic inanimate words . See intact and vocoded. Vocoding was based on the general procedures outlined in Shannon et al. [Audio stimuli were first converted to mono sound. The audio stimulus preparation produced audio stimuli at two reliability levels: n et al. . We usedn et al. , which hn et al. . To elimintact and vocoded) in terms of timing and frequency profile, in the intact condition, speech sounds were filtered below 8 kHz using a Butterworth low-pass filter (50 dB down in the stopband and 1 dB passband ripple). Furthermore, to ensure common onset timing between intact and vocoded conditions, we implemented the following steps: (1) prior to processing, zero padding of 1000 samples (21 ms) was applied to the start and the end of the original speech signal, which also eased potential filtering edge effect; (2) subsequent to envelope replacement, sine-squared ramps were applied to the zero padded segments . Note that the zero padding edge samples would always become non-zero after filtering, due to the nature of the Infinite Impulse Response (IIR). Subsequently, the amplitude envelope of the vocoded signal was replaced with that of the original speech tokens through point-by-point division and multiplication. Extraction of the envelope was as follows: first, the Hilbert envelope was computed and filtered with a 15th order Butterworth low-pass filter (cutoff at 23 Hz); second, the signal was full-wave rectified and filtered again through a 10th order low-pass Butterworth filter (cutoff at 46 Hz); finally, the signal was upshifted by a 0.005 constant value to avoid discontinuities when calculating the reciprocal of a tiny value. Replacement of envelope was done to eliminate possible confounding effects of the speech envelope due to the distortion caused by vocoding. Finally, the intact and vocoded words were normalized to the same dBA level in Adobe Audition 3.0 .To normalize the two audio conditions  using fspecial.m function. The purpose of using this blurring level was largely to preserve the global temporal cues of opening and closing the mouth while smearing spatial details necessary for specific phoneme identification, such as relative movement of the tongue relative to the teeth.The video stimuli were manipulated using in-house Matlab code. Videos of the person’s face were initially cropped to 396 × 192 pixels size, with the mouth centered and the area above the nose excluded . The deshttps://www.neurobs.com/). To mimic the size of human face, the videos were rescaled to a size of 594 × 288.Individuals sat in a sound-attenuated room, 95 cm in front of a 24-inch Dell monitor. A Vizio sound bar (model S2920W-C0) was directly placed below the monitor, with one speaker vertically aligned with the center of the monitor from which a mono sound was presented. Presentation of AV stimuli were delivered by Presentation Software  condition. One of the two blocks was animate-dominant while the other was inanimate-dominant. For half of the participants, the four animate-dominant blocks were administered before the four inanimate-dominant blocks, with a long break in between; for the other half, the order of presentation was reversed. Block order was counterbalanced using a Latin square. The dominant/oddball ratio in each block was 90:9. Based on local oddball algorithm, the number of dominant trials between two oddball trials ranged from 5 to 15, under a triangular distribution symmetrical around 10. Among the 90 dominant trials, all 40 words in that category appeared at least twice and no more than three times. The 9 words in the oddball trials were randomly selected with no repetition. In 9 out of the 90 dominant trials, a green annulus would appear after the video was launched. In this type of trial , participants were required to press “1” regardless of whether the word they heard was in the oddball category or not. These trials were later used to identify participants who did not attend to the visual stimuli and thus were excluded from further data analysis.Each trial lasted about 5.1 s, starting with a static image—the first frame of the video, which lasted for 700–1000 ms varying across trials to jitter mouth movement onset. Time of acoustic onset was further jittered due to its naturally variable latency relative to the mouth movement onset. With the duration of acoustic signals ranging from 590 ms to 1136 ms, the participants had at least 1.8 s to respond after the end of a stimulus. The final frame of the video remained on the screen until the next trial started, leaving no gap between trials. The presented words had an average sound intensity level of 54 dBA (average of slow and fast time constant) and a peak level at 73 dBA. These values were measured using a Larson Davis SoundTrack LxT1 sound level meter, based on concatenation of thirty randomly selected audio stimuli with silent gaps removed. Responses were logged by Presentation software.vocoded word pairs like “shrew” and “shoe” or “cat” and “hat”. This was rare, but noteworthy.Prior to the main testing blocks, there was a training session and a practice session. In the training session, 80 words (audio and text only) were looped in a random order and participants were asked to categorize the word by pressing “3” for animate and “4” for inanimate. This served as a means of identifying any word that looked unfamiliar or ambiguous to a participant (very rare). Feedback was provided after each trial, with debriefing after the session. The practice session used the same task as the test session. Stimuli included 10 words in each category that had been selected based on feedback from a pilot study, with both audio and video degraded. From the practice phase onward, participants were encouraged to reduce eye blinks during stimulus presentation until they responded. Participants were always required to respond to the animate words by pressing “1,” although there was no real oddball in the practice session. In total, 5 out of the 20 trials were randomly selected to serve as probe trials, as described above. During the practice session, some participants realized that it was difficult to distinguish between EEG was acquired of all subjects using 64-channel BioSemi Active Two system . EEG was acquired at a sampling rate of 1024 Hz with no high-pass filter. The EEG signal was recorded with the DC offset and drift, with an antialiasing low-pass filter (cutoff set at 1/5 of the sample rate). EEG data were down-sampled to 256 Hz using BioSemi Decimator tool after data collection. To prevent aliasing, the Decimator uses a fifth order cascaded integrator-comb filter.Response accuracy for the probe trials (72 for each person) was isolated from the analysis of other trials to assess the participant’s attention to the visual stimuli. For the remainder, besides total accuracy, each audiovisual condition was tallied respectively, collapsing across animate- and inanimate-dominant blocks. A 2 × 2 repeated measure ANOVA was conducted to see if audio and/or video degradation did substantially affect intelligibility and decision making, assuming participants did fully benefit from the training and practice sessions.clear, blurred, intact, vocoded; specific effects: clear-intact, clear-vocoded, blurred-intact, blurred-vocoded). The final mean numbers and standard deviations of trials per condition were as follows: clear-intact = 122.5 ± 21.4; blurred-intact = 121.5 ± 15.7; clear-vocoded = 119.6 ± 22.6; blurred-vocoded = 117.7 ± 21.2. These numbers increased for the main effect contrasts .EEG data were preprocessed using EEGLAB 13 . For eacStatistical analyses of ERPs between conditions were initially conducted in Fieldtrip . Given tintact vs. vocoded), the ERP waveforms of the two conditions were subjected to paired cluster-based permutation tests (CBPT). All 64 channels and sample time-points (−500 to 750 ms) were included. First, we performed two-tailed paired-samples t tests on the waveforms’ amplitude values of the two conditions for each channel and sample to determine univariate effects at the sample level. Only data samples with t values that surpassed a 0.05 probability level (two-tailed) were subsequently considered for cluster formation. Neighboring time points and channels with a univariate 0.05 p values were grouped together. The FieldTrip’s triangulation method was used to define neighboring channels. The final cluster-level statistics were calculated as the sum of all the t values within each sample-channel cluster. To evaluate the cluster-level significance, a nonparametric null distribution was created using a Monte Carlo approximation by repeating the above steps for 5000 random partitions  of the data, whereby the labels for the two conditions (samples and channels) were randomly shuffled. Following each permutation, the maximum of the cluster-level test statistics was used to create the null distribution. Significance was finally calculated by comparing the real cluster-level test statistics with the null distribution of maximum cluster-level statistics. Cluster-based differences between conditions were deemed significant if the cluster’s Monte Carlo p value was 0.05.To examine the main effects of audio or visual reliability  and as clear-intact, 97.6 ± 1.75%; blurred-intact, 97.6 ± 1.99%; clear-vocoded, 95.3% ± 2.4%; blurred-vocoded, 94.5 ± 3.25%. None of the participants reached 100% (ceiling) performance on any condition. There was an advantage of auditory reliability. Despite small percentage differences, dependent sample t-tests between conditions of the auditory reliability, with the same visual reliability condition, revealed that performance was significantly more robust for intact than vocoded stimuli regardless of the visual reliability condition . Vocoding did cause difficulty in understanding some words, which was expected since some phonemes became less distinct from others after the spectral information was smeared . The high accuracy in the different conditions as well as in the probe trials indicates that the vocoding level largely preserved intelligibility. On the other hand, there were no significant differences between the visual reliability conditions (p > 0.3).Performance (mean ± SD) was as follows: vocoded vs. intact) and visual (blurred vs. clear) reliabilities using the cluster-based permutation test (CBPT). We then test for interactions among all conditions  via post-hoc analyses of variance . The ERP value for each condition used in the ANOVA, was based on the mean ERP amplitudes within the significant time windows of the CBPTs, averaged across a frontocentral cluster of channels .We begin the ERP analyses by examining the main effects of auditory . The results show that the intact audio stimuli evoked larger N1 ERPs  and a larger subsequent broad negativity  than the vocoded audio stimuli. The t-value topographies (showing positive shifts for vocoded minus intact conditions) are consistent with typical auditory topographies  for both the N1 and the later negativity. Finally, the last cluster shows an effect (p = 0.004) that is largely reflected at parietooccipital cites , whereby a sustained negative posterior shift was observed for the vocoded vs. intact stimuli. Its topography is not consistent with the earlier auditory topographies, and thus we cannot conclude that it represents the same neural generators. We do not discuss this component further.We used the CBPT to compare the clear-intact, clear-vocoded, blurred-intact, and blurred-vocoded conditions. We subjected these values to a post hoc ANOVAs to assess for interactions. The ANOVA with the variables auditory-reliability and visual-reliability for the earlier time window (N1: 121–184 ms) revealed a main effect of auditory-reliability  = 41.3, p = 0.00001; η2 = 0.63), confirming the CBPT’s N1 effect. There was also a main effect of visual-reliability  = 6.17, p = 0.02; η2 = 0.2), which was not observed in the CBPT analysis. This is not surprising given that the post hoc ANOVA is less statistically conservative than the CBPT. Finally, there was no interaction between the variables (F = 0.45). A similar ANOVA for the later negativity (N450: 273–570 ms) revealed the main effects of auditory-reliability  = 21.3, p = 0.0001; η2 = 0.47), confirming the CBPT analysis, and visual-reliability  = 5.2, p = 0.03; η2 = 0.18)—not observed with the CBPT analysis. As a note, the visual reliability effect was observed in 19 out of 25 subjects. There was no interaction between the variables (F = 0.46).Based on the auditory-reliability CBPT we obtained the mean ERP values within the significant time windows, averaged across the nine abovementioned frontocentral cluster of channels for the clear and blurred ERPs. The left panel of p < 0.05) occurred between the conditions. The right panel shows the t-value topographies for clusters that showed significant differences (n = 3 clusters). The results show that clear vs. blurred videos evoked smaller ERPs prior to sound onset −164 to −78 ms and −54 to 12 ms; p < 0.05) and about 164–297 ms (P2: p = 0.0064) post sound onset. However, there were no cross-modal effects on the late semantic-related negativities, as we had predicted. t-Value topographies of all significant clusters were consistent with auditory generators .We used the CBPT to compare the clear-intact, clear-vocoded, blurred-intact, and blurred-vocoded, and subjected these values to a post hoc ANOVAs to test for interactions. The two ANOVAs with the variables auditory-reliability and visual-reliability for the two pre-acoustic onset periods (−164 to −78 ms and −54 to 12 ms) revealed main effects of visual-reliability  = 14.4, p = 0.0009; η2 = 0.38; 2nd period: F = 16.8, p = 0.0004; η2 = 0.41) and no effects of auditory-reliability or interaction between the variables (F < 1.5). An ANOVA for the later period (P2: 164–297 ms) revealed main effects of auditory-reliability  = 5.7, p = 0.025; η2 = 0.19) and visual-reliability  = 18.3, p = 0.003; η2 = 0.43) with no interaction between the variables (F = 0.25). The P2 auditory-reliability main effect was observed in 15 out of 25, which calls into question whether this P2 effect based on the post hoc ANOVA is reliable. Nonetheless, these results again show that effects exposed in the less conservative post hoc ANOVAs, are common to both auditory-reliability and visual-reliability contrasts. Consequently, the ANOVA’s results are not taken as confirmation of significant differences, rather they are regarded as tendencies.Based on these significant time windows, we obtained the mean ERP values at the frontocentral cluster of channels for the conditions vocoded (24-channel) audio words and blurred videos of a speaker uttering these words to assess the behavioral performance and ERPs of these conditions relative to the more reliable conditions (intact audios and clear videos). Participants performed a semantic task, whereby they identified whether the words were of animate or inanimate meaning. We hypothesized that enhanced visual reliability should result in suppression of early auditory ERPs, a process associated with enhanced AV integration efficacy [We examined the neurophysiological underpinning of speech intelligibility as a function of auditory and visual reliability. We used efficacy ,11,13,17efficacy . That isclear vs. blurred videos. This effect was not observed for the N1 ERP. The P2 has been shown to be modulated by speech-specific cross-modal inhibition, while the N1 cross-modal suppression has been associated with speech and non-speech conditions [clear videos tended to evoke larger N1s than blurred ones, as compared to smaller P2 for clear than blurred conditions. In other words, the N1 and P2 shifted in one direction. This effect is reminiscent of the findings from Shatzer et al. [clear videos evoked smaller auditory P1 and P2 ERPs but larger N1s for blurred videos. We may then acknowledge that the suppressive effect, likely instigated by the preceding visual context, is more consistent with a superimposed slow wave that begins prior to sound onset and continues past the P2 wave. The topographies of this wave suggest that it is more likely auditory in origin. If indeed AEP suppression is a consequence of a redundancy created by the preceding visual context [First, we found that the auditory P2 ERP, and ERPs occurring before sound onset, were suppressed for nditions ,29. The r et al. , whereby context , then weclear evoked larger N450 than blurred. Semantic prediction is reinforced by phonetic reliability [Second, the cross-modal effect of visual reliability on the semantic-related N450 wave was only observed in the less conservative post hoc ANOVA—iability , as demoiability ,8. Thus,Third, the cross-modal semantic prediction effect may have been weakly captured due to the high reliability of the vocoded stimuli (accuracy > 90% on average). We know from previous studies that visual context is more beneficial as acoustic degradation or noisy background increases ,2,3,30, In conclusion, the current study extends the neurophysiological cross-modal suppressive account to linguistic tasks, but also provides evidence that visual context tends to influence semantic encoding as well. Future studies addressing the current topic should consider using more degraded auditory stimuli to fully capture the semantic-related visual effect.


Inflammation plays a pivotal role in atherothrombosis. Colchicine is an anti-inflammatory drug that may attenuate this process. Cardiovascular protective effects of anti-inflammatory drugs, however, seem to be limited to patients with a biochemical response. We therefore investigated whether short-term exposure to colchicine reduced inflammatory markers and whether additional laboratory changes occur in patients with chronic coronary artery disease.In 138 consecutive patients with chronic coronary artery disease and a high sensitivity C-reactive Protein (hs-CRP) ≥ 2 mg/L, inflammatory markers, lipids, haematologic parameters and renal function were measured at baseline and after 30 days exposure to colchicine 0.5mg once daily. Hs-CRP decreased from baseline 4.40 mg/L (interquartile range [IQR] 2.83–6.99 mg/L) to 2.33 mg/L , corresponding to a median change from baseline of -40%. Interleukin-6 decreased from 2.51 ng/L (IQR 1.59–4.32 ng/L) to 2.22 ng/L , corresponding to a median change from baseline of -16%. No clinically relevant changes in lipid fractions were observed. Both leukocyte and thrombocyte count decreased (median change from baseline -7% and -4% respectively). Estimated glomerular filtration rate decreased with a mean change from baseline of -2%.In patients with chronic coronary artery disease and elevated hs-CRP, one-month exposure to colchicine 0.5 mg once daily was associated with a reduction of inflammatory markers. A small effect was seen on white blood cell count and platelet count, as well as a small decrease in estimated glomerular filtration rate. Inflammation plays a pivotal role in the complex pathophysiology of atherothrombosis ,2. Part Dampening the inflammatory response in atherosclerosis with anti-inflammatory drugs has yielded contradictory results. Anti-inflammatory treatment with canakinumab, a therapeutic monoclonal antibody targeting interleukin-1β, resulted in a detectable anti-inflammatory biochemical response and was associated with beneficial effects on major clinical outcomes in cardiovascular disease. Treatment with low-dose methotrexate however, did not show biochemical or clinical response ,8. ColchColchicine has a narrow therapeutic index. First, competition with Cytochrome P450 3A4 (CYP3A4) or P-glycoprotein metabolizing drugs may lead to decreased clearance of colchicine or alter pharmacodynamics of CYP3A4 substrates such as statins. Second, colchicine in high dose can modulate myeloid cell lines due to its anti-proliferative properties ,16. ThirThe aim of this study was to investigate whether 30 days exposure to colchicine 0.5mg once daily leads to a reduction in inflammatory biomarkers in patients with chronic coronary artery disease and to investigate effects of treatment on lipid fractions, blood indices, and renal function.st, 2017 to December 4th, 2018. Patients were eligible for participation if they had coronary artery disease proven by either invasive coronary angiography or computed tomography coronary angiography with an Agatston calcium score > = 400 units, a high sensitivity [hs]-CRP ≥2 mg/L and if they were considered clinically stable to the discretion of the caregiver. This cut-off value for hs-CRP was chosen to select patients with a pro-inflammatory status and to optimize comparability to prior clinical studies . The drug was supplied free of charge by TioPharma . The funders and drug supplier had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors confirm that all ongoing and related trials for this drug/intervention are registered and the main trial is registered in the Australian Clinical Trials Registry (ACTRN12614000093684). There are no conflicts of interests by the authors.Blood samples were available in 337 patients. Major reasons for exclusion where hs-CRP <2 (n = 184) or intolerance to the drug (n = 7), mainly in the form of gastro-intestinal upset . 138 patMedian age was 65 years and 82% were male . Of thesMedian baseline hs-CRP was 4.40 mg/L (IQR 2.83–6.99 mg/L). After one month hs-CRP was 2.33 mg/L  and after one month 2.22 ng/L , corresponding to a median change from baseline of -16% .In 57% of patients hs-CRP decrease showed concordance with IL-6 decrease. Overall, a change in hs-CRP showed a moderate correlation with change in IL-6 . The median decrease of hs-CRP and IL6 was consistent among various pro-inflammatory phenotypes such as diabetes, smoking, obesity and independent of statin use. There were no significant differences in baseline level of hs-CRP or IL-6 between these groups. .At baseline mean LDL cholesterol level was 2.38 mmol/L. No significant changes were observed in levels of LDL cholesterol, total cholesterol and triglycerides . HDL choNo change was seen in concentration of hemoglobin, hematocrit or erythrocytes . The meaThe number of leukocytes decreased from a median of 6.99 to 6.62 x 10^9/l , corresponding to a median change from baseline of -7%. The number of thrombocytes decreased from a median of 237.50 to 231.00 x 10^9/l , corresponding to a median change from baseline of -4%. Leukocyte changes were significantly correlated with changes in hs-CRP .2 at baseline and decreased to 74.99 ml/min/1.73m2 after 30 days , corresponding to a mean change from baseline of -2%. Mild or moderate renal impairment (eGFR 30 to 60 ml/min/1.73m2) was seen in 11% of patients at baseline and 14% of patients at follow-up. Change in eGFR did not correlate significantly with the change in hs-CRP .Creatinine increased from a mean of 87.90 to 89.53 μmol/L , corresponding to a mean change from baseline of 1% . DerivedThis study showed that patients with chronic coronary artery disease and a pro-inflammatory state defined by hs-CRP ≥ 2 mg/L, had a significant decrease in both hs-CRP and IL-6 after one month of colchicine exposure. No clinically relevant changes in lipid spectrum were observed. A small decrease in mean corpuscular volume was observed, as well as a decrease in leukocyte and thrombocyte number and a small decrease in estimated glomerular filtration rate. The present study extends our knowledge of the anti-inflammatory effect of 0.5mg colchicine once daily to patients with chronic coronary artery disease.The direction and extent of the change in inflammatory markers after exposure to colchicine in this pro-inflammatory population with coronary artery disease compares to observations from previous studies in heart failure and patients with metabolic syndrome. Although absolute reduction of hs-CRP and IL-6 were larger in these studies in which baseline levels were higher, the relative reduction was similar. The reduction in the current study was achieved with a lower dose of colchicine ,22. SystThe magnitude of hs-CRP and IL-6 reduction found in the current study is similar to the effect of canakinumab used in the Canakinumab Antiinflammatory Thrombosis Outcome Study (CANTOS) . AlthougIn contrast to hs-CRP, the evidence for a causal relation to levels of IL-6 and in particular IL– 1 beta and atherothrombosis has become more rigid after observational Mendelian randomization studies and results of the CANTOS trial ,28–30. AIL– 1 beta is an upstream biomarker of IL-6 and there are data suggesting that high doses of colchicine can reduce IL– 1 beta release. However, such an effect has not been described in patients with chronic coronary artery disease . The absA small reduction in leukocyte and thrombocyte count was observed during one month of colchicine exposure. This finding is not unexpected. Colchicine affects leukocyte adhesion and chemotaxis and also has an anti-proliferative effect on leukocytes, mediated by the ability to disrupt the cytoskeleton. Colchicine irreversibly binds with tubulin to form an intracellular tubulin complex preventing the formation of microtubules ,34. In hA small increase in serum creatinine and a corresponding decrease of eGFR was observed. Excretion of colchicine takes place mainly via the enterohepatic circulation (80%) and in part via renal excretion (20%) ,36. WhetA methodological limitation of our study is the paired testing with the absence of a parallel control group. This is inherent to the design of the study as it was part of the run-in phase prior to randomization of three sites in a clinical trial. Due to the selection of hs-CRP ≥2, part of the change in hs-CRP may be explained by regression to the mean or natural course. However, changes in hs-CRP are more pronounced than those observed in the placebo arm of the similar CANTOS population (change of -40% in this cohort versus -17% in CANTOS), suggesting the CRP decrease is perhaps only partially influenced by preselection and natural course. In addition, no change in IL-6 was seen in the placebo arm of CANTOS [Finally, the subgroup analyses have an exploratory purpose only. Since the study was not designed to assess differences between subgroups, the limited sample size increases risk for type 2 errors in these observations.One-month exposure to low-dose colchicine was associated with a reduction of the inflammatory markers hs-CRP and IL-6 in patients with chronic coronary artery disease and baseline hs-CRP ≥2 mg/L. An effect was seen on estimated glomerular filtration rate, leukocyte and thrombocyte cell count, which warrants placebo-controlled and longer follow-up of these parameters. Whether the anti-inflammatory and other effects observed in this study are solely contributable to colchicine should be confirmed in placebo-controlled assessment. Whether this effect translates into a clinical benefit has to be awaited. Observations thus far support ongoing clinical research in colchicine as anti-inflammatory drug in atherosclerosis.S1 Checklist(DOC)Click here for additional data file.S1 File(DOCX)Click here for additional data file.


Valid instruments for measuring physical activity at the low end of the physical activity range and producing quantitative results are required among dialysis patients who are extremely inactive. This study aimed to translate and adapt a Chinese version of the low physical activity questionnaire (LoPAQ) and to examine its reliability and validity among hemodialysis patients.This was a cross-sectional study. The LoPAQ was translated into Chinese and culturally adapted following the standardized questionnaire adaptation process. Participants wore an ActiGraph for seven consecutive days and were asked to complete the Chinese version of the LoPAQ (C-LoPAQ) following the ActiGraph monitoring period. The criterion validity of the C-LoPAQ was examined with accelerometers using Spearman’s correlation coefficients. Bland-Altman plots were adopted to determine the absolute agreement between methods. The test-retest reliability was analyzed using the intraclass correlation coefficient (ICC).rho = 0.47, p < 0.01). A moderate correlation was also observed between the C-LoPAQ and the ActiGraph-measured physical activity for total calories . There was a fair correlation between ActiGraph-measured sedentary time and C-LoPAQ-measured inactive time . The test-retest reliability coefficients of C-LoPAQ ranged from 0.30 to 0.66.Eighty-five hemodialysis patients had valid accelerometers and C-LoPAQ data. The total walking time reported on LoPAQ was correlated with step counts by ActiGraph (The C-LoPAQ demonstrated moderate validity for measuring low levels of physical activity, especially walking, and total kilocalories of physical activity among hemodialysis patients in China.The online version contains supplementary material available at 10.1186/s12882-021-02230-3. Chronic kidney disease (CKD) has become a public health issue, affecting approximately 8–16% of the adult population worldwide . With irIn MHD patients, a low physical activity level is closely related to poor prognosis . SeveralPhysical activity refers to all skeletal muscle movements that result in energy consumption. It can be classified into low, moderate, and high levels according to energy consumption , 13. PatThe common questionnaires for evaluating physical activity include the 7-day Physical Activity Recall Questionnaire (PAR), Human Activity Profile (HAP), and International Physical Activity Questionnaire (IPAQ), which have been used in CKD patients in previous studies , 16, 18.The low physical activity questionnaire (LoPAQ) developed by Johansen et al.  emphasizThis was a cross-sectional study. Patients undergoing hemodialysis treatment were recruited from outpatient hemodialysis units of two hospitals  in mainland China through convenience sampling from December 2018 to March 2019. The inclusion criteria were as follows: 1) age ≥18 years; 2) on hemodialysis for ≥3 months; 3) able to walk without assistance; and 4) ability to provide informed consent and complete the questionnaires. The exclusion criteria were as follows: 1) diagnosis of mental or cognitive disorders; 2) unstable conditions; and 3) hospitalization in the previous 3 months. The study was approved by the Human Subjects Ethical Sub-committee of the Shanghai Jiao Tong University (SJUPN-201705). Written informed consent was obtained from all participants prior to data collection.rho = 0.62, p < 0.001), the Physical Function score of the 36-item Short Form Health Survey , and physical performance indexes [The original English version of the LoPAQ was developed by Johansen et al. . It is a small, watch-like, unobstrusive device that measures acceleration in three axes. The device was initialized using 60-s epochs to collect data at a sampling rate of 30 Hz. Participants were instructed to wear the ActiGraph on the right hip on an elastic waistband during waking hours, except when bathing, swimming, or doing other water activities, for seven consecutive days. On the last data collection day, a research assistant went to the dialysis unit to remove the accelerometer. Raw accelerometer data were downloaded using ActiLife 6 software (ActiGraph) and were analyzed in 60-s epochs. The default filter settings were adopted. Wear time was validated using a filter of 150 consecutive zero-count minutes, with allowance for < 1 min of activity counts < 100 counts per minute . At leasA questionnaire on the demographic and clinical information of the patients, consisting of two parts, was self-developed. Part I included demographic information such as age, sex, education, marital status, residence, and living conditions. Part II recorded the clinical data, including the cause of chronic renal failure and dialysis vintage.After obtaining written consent to participate in the study, each participant completed a demographic questionnaire during the dialysis session and received an ActiGraph for 7 days. After 7 days, the research assistant visited the dialysis center again to retrieve the ActiGraph. On the ActiGraph removal day, the research assistant administered the C-LoPAQ face-to-face. The participants were asked to recall their physical activity and sedentary behavior over the past 7 days for the same time the ActiGraph was worn. All clinical information was extracted from the inpatient information system of the hospital. After two weeks, the participants were invited to complete the C-LoPAQ again to evaluate test-retest reliability.p < 0.05 was considered statistically significant.Statistical analysis was performed using SPSS 21.0 software . Demographic information was presented using descriptive statistics, including mean, standard deviation (SD), and percentage. The levels of skewness and kurtosis were determined to assess the normality of each variable . ContentA total of 96 participants were recruited for the validity test. A total of 85 MHD patients who had valid ActiGraph and C-LoPAQ data were included in the data analysis, among whom 56 (65.9%) were from Renji Hospital  and 29 (34.1%) were from Tongren Hospital. The mean age of the participants was 62.3 (SD 11.8) years, and 57% were male. The majority of the participants was married (85%) and retired (82%). The most common cause of renal failure was glomerulonephritis, followed by diabetic nephropathy. The average duration of dialysis was 4.9 (SD 3.7) years  steps/day, and 72.9% of the participants had an average step count of less than 5000 steps/day. In addition, 83.7% of the activity was light-intensity physical activity on average. The average energy expenditure was 986.1 (696.6) kcal/week. For the C-LoPAQ, the mean walk time was 28.4 (27.5) min/day. The total energy expended from walking and other activities was 1170.9 (820.1) kcal/week. Participants reported a mean sitting time of 4.7 h ” as examples of “light activity” in the original English version of the LoPAQ was not appropriate, as Chinese MHD people do not usually engage in those exercises. Therefore, they were removed. Examples such as “light yard or gardening work” and “chair exercise” were replaced by “watering the plants” and “walking downstairs.” In the subscale of “moderate activity,” “aerobics class” and “swimming (the side stroke or breast stroke)” were replaced by “TaiChi” and “square dancing,” and “softball” and “downhill skiing” were deleted. In the subscale of “vigorous activity,” “playing tennis or racquetball” and “cross-country skiing” were removed and replaced with “slow rope skipping.” After the revisions, the panel rated the content validity of the C-LoPAQ using a 4-point Likert scale. The S-CVI was 0.91, and the CVR was 0.98.rho = 0.44, p < 0.01). The total walking time reported on C-LoPAQ was also correlated with step counts by ActiGraph . There was a fair correlation between ActiGraph-measured sedentary time and C-LoPAQ-measured inactive time   Table .Table 3The results from the Bland-Altman plot show a mean difference of 184.8 kcal of energy expenditure per week between C-LoPAQ and ActiGraph data. In addition, MHD patients who were relatively active tended to over-report physical activity using the C-LoPAQ, and an error can be seen from over-reporting as the mean increases. The limits of agreement for total calories between the C-LoPAQ and ActiGraph had wide ranges, from − 1410.0 to 1779.6, with 6 outliers (7.1%)  at the first test and 1732.7 SD 1404.5) kcal/week at retest. For test-retest reliability, the ICCs ranged from 0.301 to 0.663 for the subscale scores. The details are shown in Table 04.5 kcalThe aim of this study was to develop the C-LoPAQ and to determine its reliability and validity for measuring low levels of physical activity, especially walking, total kilocalories of physical activity, and time spent sitting compared with ActiGraph data from MHD patients. The main findings indicated that the walking times obtained by the C-LoPAQ moderately correlated with step counts measured by the ActiGraph. The total energy expenditure reported by the C-LoPAQ also correlated well with the kilocalories obtained from the ActiGraph. The C-LoPAQ-measured inactive time was associated with the ActiGraph-measured sedentary time. However, the C-LoPAQ had a fair test-retest reliability.In the present study, 72.9% of MHD patients were sedentary (< 5000 steps/day) with a mean of 3807.8 (2723.0) steps/day. This finding is similar to a previous multicenter study in China, in which a mean daily step count of 3759.9 (2664.5) was documented among 320 MHD patients  using pedometers . The levIt is important to assess the types of physical activities that are commonly engaged by the study population in a certain cultural context, as it is an indispensable part of content validity. Several examples of different intensities of physical activities were replaced with activities considered more culturally appropriate for Chinese dialysis patients’ lifestyles, or they were deleted altogether. For example, “aerobics class” was replaced by “TaiChi,” and “playing tennis or racquetball” was replaced by “slow rope skipping.” These replacements were achieved using activities with similar energy expenditure based on the Compendium of Physical Activities , 35. Therho = 0.47) that was very close to that reported in a previous study [rho = 0.44) in measurements of total energy expenditure between the C-LoPAQ and accelerometer data. This result is relatively lower than that of a previous study, in which a correlation coefficient of 0.58 was found between the energy expenditures during walking measured by the LoPAQ and the Minnesota Leisure Time Activity Questionnaire [The C-LoPAQ showed reasonable evidence of validity for walking time, as it was moderately correlated with ActiGraph-measured step counts, with a Spearman correlation . However, the correlation did not reach the cut-off point (above 0.4) for what was generally considered acceptable validity. Such a low correlation may be due to different interpretations of sitting behavior among respondents. Some participants may not count lying down while not reading or watching television during waking time as sitting behavior. As suggested by Heesch et al. [Results from the current study demonstrated fair validity of the C-LoPAQ for sedentary behavior. Hours of sitting activity on the C-LoPAQ positively correlated with ActiGraph-measured sedentary time  for walking time, compared to moderate and vigorous intensity activities, may be attributed to variability in walking behaviors in our study. In order to minimize the bias associated with learning or carryover effects from previous responses , a two-wA number of limitations should be addressed. First, even though the ActiGraph provides objective and valid measures of physical activity and sedentary behavior, it is not able to capture information on ambulatory activities, such as cycling, weightlifting, or water-based activities, such as swimming. This may overestimate sedentary behaviors. Second, a period of 2 weeks between the first and second tests may significantly affect the results of the reliability of the C-LoPAQ. Third, responsiveness or sensitivity to detect meaningful changes in the C-LoPAQ was not determined in the current study. Finally, as the size of the sample is relatively small, its generalizability might be an issue.The C-LoPAQ is easy to use and demonstrated an acceptable validity for measuring low levels of physical activity, especially walking, and total kilocalories of physical activity among dialysis patients in China. As a relatively low correlation was observed between sitting time in the C-LoPAQ and the ActiGraph-measured sedentary time, further clarification on the meaning of sitting time in the C-LoPAQ should be provided. Because the amount of physical activity may vary, re-testing of the C-LoPAQ should cover the same time period, consequently improving the accuracy of test-retest reliability. Further studies are needed to determine the responsiveness of the C-LoPAQ.Additional file 1.


Anal squamous cell carcinoma (SCC) is a rare cancer often caused by infection with high-risk human papillomavirus (HPV). Immune checkpoint blockade (ICB) targeting PD-1 or PD-L1 is successfully used for the treatment of other cancers as melanoma and lung cancer and might be an option for anal SCC as well. PD-L1 expression is associated with ICB outcome for example in melanoma. Here, we investigated whether infection with HPV affects PD-L1 expression and observed that almost two-thirds of anal SCC patients had PD-L1 positive tumors. Interestingly, these patients had a better median overall survival (OS). Sex, grade of differentiation, and HPV infection status did not influence the median OS. HPV infection status and PD-L1 expression were not correlated. Our findings indicate that PD-L1 is an independent prognostic marker in anal SCC. Besides, ICB targeting PD-1 or PD-L1 might be a therapy option for anal SCC patients irrespective of HPV infection status.n = 39, 72.2%). Four patients were infected with multiple HPV subtypes. HPV infection was significantly more often detected in female than in male patients . Patients with PD-L1 positive tumors showed a significantly better median overall survival (OS) compared with patients with PD-L1 negative tumors . The median OS was significantly different among the distinct tumor stages (p = 0.029). Sex, grade of differentiation, and HPV infection status did not influence the median OS. Furthermore, HPV infection status and PD-L1 expression were not correlated. A multivariate Cox regression analysis revealed that PD-L1 expression status was an independent prognostic marker for survival (p = 0.012).Anal squamous cell carcinoma (SCC) is a rare cancer with increasing incidence. Infection with high-risk human papillomavirus (HPV) subtypes is the major cause for its development. We retrospectively analyzed tumor samples from 54 anal SCC patients for infection with a panel of 32 HPV subtypes in a PCR-based approach, determined the PD-L1 expression status, and correlated the findings with the clinical data and the survival of the patients. Forty-two patients (77.8%) were HPV-positive and harbored at least one carcinogenic HPV subtype. HPV16 was the most frequently detected ( Anal cancer is a rare cancer entity, affecting about 40,000 patients worldwide . HoweverStandard of care for most anal cancer patients is still a chemoradiotherapy (CRT) scheme, originally described by Nigro in the 1970s, that combines radiation therapy and chemotherapy with 5-fluorouracil and mitomycin . FurtherThe introduction of targeted therapies and immune checkpoint blockade (ICB) has tremendously changed the field of oncology in the last 10 years. ICB targets immune checkpoints, such as the CTLA-4 or the PD-1/PD-L1 axis with specific antibodies, and has achieved significant results in several cancer entities, including melanoma, non-small cell lung cancer (NSCLC), and squamous cell carcinoma of the head and neck (HNSCC) ,8,9. RecIn this study, we analyzed tumor samples from 54 anal SCC patients for infection with a panel of 32 distinct HPV subtypes, determined the PD-L1 expression status on the tumor cells, and correlated the findings with patient survival in order to elucidate the relationship between infection with different HPV subtypes and PD-L1 expression in anal SCC.The baseline characteristics of the study population are summarized in A total of 54 patients with anal SCC from stage I to stage IV were included in the study—31 of them were female (57.4%). Eight patients were categorized as stage I, 23 as stage II, 12 as stage III, and 11 as stage IV anal SCC. The median age at diagnosis was 60.1 years, ranging from 28.5 to 81.7 years. In most cases, the tumors were located at the anal canal and anal sphincter , followed by perianal skin . The majority of tumors were poorly differentiated (51.9%).INK4A (p16) [The viral oncoproteins E6 and E7 significantly contribute to HPV-induced carcinogenesis by deregulating the expression and function of several cell cycle-regulating proteins, including p53, pRb, and p164A (p16) . Hence, 4A (p16) ,18. In op = 0.018) and three of the four patients infected with multiple HPV subtypes were women.HPV DNA was detected in 42 out of 54 samples 77.8%, . All HPVThe PD-L1 expression status was evaluable in the tumor samples from 53 patients, and the staining intensity was evaluated and graded according to the tumor proportional score (TPS), as HPV is epitheliotropic .p = 0.019).The PD-L1 expression was detectable (TPS ≥ 1%) in 33 tumor samples (61.1%). An extremely high PD-L1 expression  was observed in four samples (7.4 %). Notably, three of them were HPV-negative. In 20 samples (37.0%), no PD-L1 expression was detectable . Interestingly, a PD-L1 expression ≥ 10% (TPS category 3 and higher) was significantly more often detected in men  than in women  or even absent  in the tumor samples of the four patients infected with multiple HPV subtypes. There was no significant difference of HPV infection status in tumors with vs. those without PD-L1 expression (p = 0.7285).Comparing the HPV status and PD-L1 expression revealed that 27 out of 42 HPV-positive patients (64.3%), and 6 out of 11 HPV-negative patients (54.5%) showed an expression of PD-L1 . The PD-p = 0.006). As expected, the median OS was significantly different among the distinct tumor stages (p = 0.029). Further subgroup analyses showed that the median OS was 68.3 months for men (n = 23) and 102.5 months for women (n = 31), but the differences were not statistically significant (p = 0.906). Additionally, no statistically significant survival differences were observed for good and intermediate vs. poorly differentiated tumors (p = 0.274). There was no difference of OS in patients with HPV-negative (n = 12) vs. those with HPV-positive (n = 42) tumors . Neither was there an OS difference between patients with HPV16-positive and those with HPV-negative tumors (p = 0.189). Due to the small number of cases, we did not analyze the survival of patients infected with single vs. multiple HPV subtypes. Furthermore, there were no statistically significant differences in OS between patients with ≥10% PD-L1-expressing tumor cells (TPS category 3 or higher) and patients with <10% PD-L1-expressing tumor cells , between people living with HIV and HIV-negative patients (p = 0.714), and between immunosuppressed and immunocompetent patients (p = 0.948). The median OS of all the patients was 68.3 months. The one-year, five-year, and ten-year OS rates were 91%, 55%, and 29%, respectively . The subp = 0.012). In this model, the tumor stage  and HPV16 infection status  were not associated with patient survival.In a multivariate cox regression model, PD-L1 expression was an independent risk factor and was associated with a favorable OS , a multicenter open-label phase Ib study, investigated the safety and efficacy of the anti-PD-1 antibody pembrolizumab in various cancer entities, including anal SCC . PatientPD-L1 was also examined in anal cancer, besides its putative role as a biomarker for predicting immunotherapy response. Evidence concerning PD-L1 expression and survival outcome for patients receiving CRT is controversial. Govindarajan et al. observed a poorer outcome of PD-L1 positive patients , and anoHPV infection is a major risk factor for developing anal SCC . Our stuIt would be interesting to investigate if the survival of the patients and the PD-L1 expression of their tumors differed between tumors with different HPV subtypes. However, the number of the patients infected with subtypes other than HPV16 was too low for separate statistical analyses of the distinct subgroups. In our patient cohort, 90.3% of women, but only 60.9% of men, were infected with HPV. Interestingly, a small subgroup of four patients (7.4%) was infected with multiple HPV subtypes. This phenomenon has been previously observed in the cervical cells of HPV-positive women , but alsExcept for the PD-L1 expression status, we did not find any significant correlations of sex, differentiation, or HPV infection status with the survival of the patients. This was quite surprising, as an improved OS for HPV-positive patients was observed in previous reports ,32. HoweVaccination against HPV might be able to boost the intrinsic anti-tumor response by reactivating immune cells to recognize and destroy tumor cells presenting viral antigens. Furthermore, it might also alter the expression of immune checkpoints on tumor cells or immune cells. Today, HPV infection with the two most common subtypes, HPV16 and HPV18, can be effectively prevented by the three commercially available vaccines, which cover distinct HPV subtypes that are associated with HPV-related cancers and the formation of genital warts ,34,35. HThis study has some limitations. Anal cancer is a rare cancer entity and the availability of tumor material is sparse, thus we were not able to include a consecutive series of patients in this analysis. Furthermore, because of the exploratory nature of this study and the limited number of available samples, we did not perform a hypothesis-driven sample size calculation. The number of included patients was certainly small and the study may lack the power to detect significant differences, in particular regarding the subgroups of HPV-positive and HPV-negative patients. Furthermore, we could not conclude whether or not PD-L1 expression was linked to a response to immune checkpoint blockade, as the population had not been treated with immunotherapy. Nevertheless, it is possible that PD-L1 expression acts as prognostic factor, regardless of treatment with immune checkpoint blockers. Tumor stage was associated with the OS in the univariate analysis, but not in the multivariate Cox regression model. We assumed that this was due to the small number of patients in the stage subgroups and the uneven distribution of the patients.The study was a retrospective analysis of 54 patients who were diagnosed and treated for anal SCC at the LMU University Hospital Munich between 15 February 1998 and 13 September 2017. Patients were included when archived tumor samples were available for further analyses. All of the patients received stage-adapted standard of care treatment, irrespective of HPV expression . The clinical data and treatment outcomes were extracted from the patient records and were merged into a central database. The institutional review board of the medical faculty of the University Hospital approved the study . Because of the explorative nature of this study, no power calculations were made in advance and the population was sampled for the availability of tissue specimens.n = 53). In one case, the PD-L1 and p16 expression analyses were not possible because of technical reasons. The sections were derived from surgical excisions, while biopsies were excluded and not assessed. Sections from entire tumor blocks were used to account for the intratumoral heterogeneity and variability of the PD-L1 expression. They were processed as described previously [® microscopy slides . Heat-mediated antigen retrieval was performed with a pressure cooker on a Ventana Benchmark XT autostainer  with the XT UltraView diaminobenzidine kit , and counterstained with hematoxylin (Vector Laboratories). Immunohistochemistry was performed for PD-L1 with clone SP263  and for p16 with clone E6H4E6H4/p16INK4a . The percentage of PD-L1 positive cancer cells was independently evaluated by two trained dermato-pathologists (Markus V. Heppt and Markus Reinholz) and categorized with the tumor proportional score (TPS), as previously described by Scheel et al. [The PD-L1 and p16 expression of the tumor cells was determined by immunohistochemical staining in archived formalin-fixed paraffin-embedded (FFPE) tissue sections  were examined under an upright microscope and compared with the corresponding hematoxylin and eosin-stained sections. The representative tumor areas were marked manually by a trained dermato-pathologist (Markus V. Heppt). Subsequently, the sections were rehydrated with PCR-grade H2O, a 5 × 5 mm part of the tumor tissue was transferred into a tube containing 20 µL extraction buffer  and was digested over night at 56°C. Afterwards, the samples were incubated for 10 min at 95 °C to inactivate proteinase K, and the HPV DNA was amplified using the polymerase chain reaction (PCR)-based LCD Array HPVType 3.5 , according to the manufacturer’s instructions with minor modifications. The PCR included an initial denaturation step , followed by 45 cycles  and a final elongation step . Hybridizing of the amplified samples with the array plates was conducted according to the manufacturer’s instructions. For HPV subtype identification, the LCD array plates were scanned in a CHIP Scanner PF7250 (Chipron) and analyzed using SlideReader V12 Software (Chipron). The results were confirmed by examining the slides with a microscope at 40× magnification.FFPE tissue sections .Overall survival (OS) was defined as the time from initial diagnosis until death due to any cause. Survivors were censored at the time of the last documented follow-up. The survival probabilities were calculated with the Kaplan–Meier method and the survival curves were compared with the log-rank test. To investigate the relationship of the factors of interest with OS, a multivariate Cox proportional hazards regression model was calculated, as performed previously ,40. HazaIn conclusion, our findings indicate that PD-L1 is expressed in both HPV-positive and -negative anal SCC, irrespective of the HPV subtype, indicating that ICB may be a promising treatment option for this patient group.


The evaluation of the response to vaccination in patients with inborn errors of immunity is a tool to evaluate T-dependent and T-independent antibody residual function of B lymphocytes and it is part of the diagnostic definition for Common Variable Immune Deficiencies. Currently used classifications for Common Variable Immune Deficiencies patients are based on the frequency of B cell subsets, and have been proven as a valid instrument for identification of patients at higher risk of infectious and non-infectious complications. This 6-years period observational study delineated the measurement of specific IgA antibodies induced by a 23-valent pneumococcal polysaccharides vaccine by a standardized ELISA for the quantification of IgA antibodies to all 23 pneumococcal serotypes as an additional prognostic marker in 74 CVID patients. The inability to mount an IgA-mediated response against the pneumococcal polysaccharide antigens or the inability to maintain the antibody response over time identified poor IgA CVID responders with severe immunological impairment, great risk of co-morbidities, and poor prognosis. The division of CVID patient into specific IgA-non responders and IgA-responders discriminated better than other CVID classifications for infectious risk, while it overlapped for non-infectious complications. Our study suggested to add the evaluation of the antibody response by the 23-valent IgA assay in the clinical monitoring of CVID patients. Immunoglobulin serum levels and antigen-specific antibodies are included as diagnostic criteria in the assessment of patient with inborn defects of immunity . A conseCommonly used classifications for CVID patients are based on the frequency of B cell subsets. These immune-phenotypic classifications, and in particular the studies done by EUROclass group  and by tWe conducted an observational study over a 6 years period in a population of CVID patients immunized by Pneumovax® to define if IgA-mediated responses to pneumococcal polysaccharides could be an additional prognostic marker in CVID patients. The results allowed to identify CVID patients with a more severe immunological impairment, a greater risk of co-morbidity, and a worse prognosis. The IgA response discriminates better than other classifications for infectious risk, while it overlapped for non-infectious complications.This single-center study was carried out in the Referral Center for Adult Primary Immune Deficiencies of Rome, Italy. Seventy-four CVID patients  and 20 healthy volunteers (HD)  were enrolled. All patients included in this prospective observational study over a 6-years period were diagnosed as CVID following the ESID/PAGID criteria . In partPatients and HD were immunized with 1 dose of Pneumovax®  a 23-valent polysaccharide vaccine which contains the pneumococcal serotypes 1, 2, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20, 22F, 23,F and 33F. Based on our previous study , Birmingham, UK), with pre-coated microtiter plates with all 23 pneumococcal polysaccharide serotypes present in the Pneumovax vaccine. Absorption of interfering anti-cell wall polysaccharide (CPS) antibodies was incorporated in this assay. In a previous paper  we have Flow Cytometry Analysis of peripheral blood mononuclear cells was performed with the combinations of 4 fluorochrome-labeled monoclonal antibodies, all obtained from BD Biosciences. The B-cell populations were identified as classical naïve (CD19+CD27-CD21+CD38+), switched memory (CD19+CD27+CD21+IgM–), IgM memory (CD19+CD27+IgM+IgD+), transitional (CD19+IgM++CD38++), CD21low  using the CellQuest (BD) and FlowJo (Tree Star) software as we previously described . On the (SD) where appropriate. Comparisons of continuous parameters between treatment groups were calculated with a t-test if normally distributed and with a Mann-Whitney U-test if not normally distributed; differences in frequencies between groups were calculated by using the χ2 exact test. Time to event was calculated by Kaplan-Meier curves and expressed elapsed time to baseline Kaplan-Meier curves for CVIDs were split into two groups: (1) IgA responders; (2) IgA non-responders. The statistical significance was set at the conventional level of P < 0.050. All statistical analyses were performed using the statistical package Stata 11  and GraphPad7 .Patient demographics and clinical characteristics are summarized by frequencies and percentages and means and standard deviations Baseline characteristics of the 74 enrolled CVID patients are summarized in The frequency of total B cells was similar within the groups. Patients from the IgA-NR group had an increased frequency of naïve B cells and a lower frequency of switched memory B cells compared to the IgA-R. The frequencies of marginal zone B cells, transitional B cells and CD21low B did not differ in the two groups . Frequenp = 0.003; HR 2.85, 95% CI 1.4–5.7, p = 0.013). We observed a similar number of episodes/year in IA group and in group IB, and a lower number of episodes in group II . Likewise, a similar number of episodes/year were observed in IA group (0.7 ± 1) and IB group (0.5 ± 0.7) and a lower number in II group (0.2 ± 0.4). CVID patients are also prone to develop non-infectious complications. The prevalence of bronchiectasis was 53% in IgA-NR vs. 14% in IgA-R  for CVID participants was 64 ± 18.5 months. IgA-NR had a 2.8-fold higher risk to develop URTI in comparison to IgA-R (Log-rank p < 0.02).At the second assessment (post 2), 36 ± 6 months after immunization, among IgA-R patients we identified a subgroup of five patients that showed a long lasting response with a level of specific anti PnPS IgA above the identified cut off (175 ± 13 U/ml) while 4 patients lost the specific IgA (19 ± 18 U/ml) . In the In clinical practice, in the evaluation of the ability to respond to immunization induced by pneumococcal polysaccharide vaccine or polysaccharide-conjugated pneumococcal vaccine, the dosage of IgG specific anti-PnPS pre- and post-vaccination is usually used. The WHO has recommended considering an IgG cut-off ≥0.35 micrograms/ml of pneumococcal anti-serotype to each polysaccharide tested to evaluate the capacity of pneumococcal vaccines to give a post-vaccine protective response . The evaIn the immunocompetent subjects, for the measurement of the anti-PnPS IgA antibodies there are few data available in the literature concerning the reference values, and therefore the interpretation of the data on the efficacy of the antibody response can be difficult –22. It hIn this study, we defined how the measurement of post-vaccine anti-PnPS IgA levels is a diagnostic parameter capable of classifying patients into groups with different risk of developing CVID-related complications over time. In particular, in the 6-years longitudinal study, we have evaluated the potential predictive value of the residual capacity to mount a specific IgA response. The inability to mount an IgA-mediated response against the pneumococcal polysaccharide antigens or the inability to maintain the antibody response over time was associated to a greater frequency of URTI and LRTI exacerbations due to a variety of pathogens , 25 duriIn the classification of inborn errors of immunity, CVID is considered an heterogeneous clinical and immunological condition characterized by antibody defects . The widIn conclusion, the vaccine response to pneumococcal polysaccharide antigens by the standardized 23-valent IgA assay is easy to interpret. The evaluation of anti-PnPS23 IgA has been shown to be a prognostic marker, allowing to identify good and poor CVID responders. We suggest to add this test to identify patients with serious immunological impairment, a greater risk of co-morbidity and a worse prognosis who could benefit from closer clinical monitoring, and from additional preventive measures, including antibiotic prophylaxis . We suggThe datasets generated for this study are available on request to the corresponding author.The studies involving human participants were reviewed and approved by Ethical Committee of Sapienza University. The patients/participants provided their written informed consent to participate in this study.IQ, FP, and CM conceptualized the study. IQ, FP, CM, IM, and FC designed the protocol study. FMC performed all laboratory tests. FP, CM, IM, and FC recruited patients and collected data. FP and FC did the statistical analysis. IQ, FP, CM, and IM prepared the first draft of the manuscript. All authors reviewed the manuscript before publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


A receptor (GABAAR) agonist, were microinfused into the rat BLA before or after BLA priming. Our findings show that BLA GABAAR activation via muscimol mimicked the previously reported effects of electrical BLA priming on LTP in the perforant path and the ventral hippocampal commissure-CA1 pathways, decreasing CA1 LTP and increasing DG LTP. Furthermore, muscimol application before or after tetanic stimulation of the ventral hippocampal commissure-CA1 pathways attenuated the BLA priming-induced decrease in CA1 LTP. In contrast, muscimol application after tetanic stimulation of the perforant path attenuated the BLA priming-induced increase in DG LTP. The data indicate that GABAAR activation mediates metaplastic effects of the BLA on plasticity in the CA1 and the DG, but that the same GABAAR activation induces an intra-BLA form of metaplasticity, which alters the way BLA priming may modulate plasticity in other brain regions. These results emphasize the need for developing a dynamic model of BLA modulation of plasticity, a model that may better capture processes underlying memory alterations associated with emotional arousing or stressful events.The term “metaplasticity” is used to describe changes in synaptic plasticity sensitivity following an electrical, biochemical, or behavioral priming stimulus. For example, priming the basolateral amygdala (BLA) enhances long-term potentiation (LTP) in the dentate gyrus (DG) but decreases LTP in the CA1. However, the mechanisms underlying these metaplastic effects are only partly understood. Here, we examined whether the mechanism underlying these effects of BLA priming involves intra-BLA GABAergic neurotransmission. Low doses of muscimol, a GABA The term “metaplasticity”  is used Stress-related alterations of memory are mediated by amygdala–hippocampus interactions, as demonstrated in many human and animal studies ,16,17,18Specifically, in previous studies, we showed that, while activation of the BLA enhances LTP at hippocampal perforant path-dentate gyrus (DG) synapses, it impairs LTP in the ventral hippocampal commissure-CA1 pathway (vHC-CA1) ,26. ThesA receptors in the BLA has been suggested to play a critical role in the control of synaptic transmission and plasticity and thereby in stress-induced modulation of memory [One such alternative mechanism may involve gamma-aminobutyric acid (GABA) neurotransmission, since studies have demonstrated that the BLA contains a powerful inhibitory GABAergic circuit ,29. The f memory ,33,34,35A receptors in this brain region, as well as the role of the GABAA receptors in memory and anxiety states [Considering the anatomy and the function of the BLA, the distribution of the GABAy states ,37, GABAA receptor activation mediates BLA metaplastic effects on hippocampal plasticity. However, such activation induces intra-BLA form of metaplasticity, which affects the very same BLA-induced metaplasticity in the hippocampus. The results call for the development of a dynamic description of BLA effects on plasticity in other brain areas, a concept that will reflect on developing translational intervention approaches in stress and trauma-related disorders.The primary purpose of this study was to first examine the involvement of intra-BLA GABAergic neurotransmission in the metaplastic effects of the BLA on hippocampal synaptic plasticity. Secondly, it was to test whether priming the BLA before or after this manipulation leads to intra-BLA metaplastic effects. We demonstrated that, indeed, intra-BLA GABAMUS groups  were not significantly different from the CONTROL groups . In contrast, microinjection of muscimol into the BLA induced a significant increase in baseline excitation in the PP-DG pathway  = 13.85; p < 0.001; CONT-VEH and BLA-VEH) and MUSCIMOL groups (CONT-MUS and MUS-BLA) one to ten minutes post drug injection .To gain insight into GABAergic regulation of BLA modulation of synaptic plasticity in the DG and the CA1, muscimol was microinfused into the BLA, either as a standalone manipulation or 10 min before or after priming of the BLA prior to LTP induction in the hippocampus  ,39. MuscCONT-VEH: 314.2 ± 22.85%; CONT-MUS: 167.25 ± 26.86%; CONT-MUS-post: 202.48 ± 24.29%; p < 0.0001), with a significant difference between groups  = 11.7; p < 0.001) and groups–time interaction  = 18.72, p < 0.0001). Fisher post hoc test revealed significant differences between both CONT-MUS and CONT-MUS-post groups vs. CONT-VEH group (p < 0.0001 for both) and CONT-MUS group vs. CONT-MUS-post group (p = 0.03).Next, we examined the effect of manipulating GABAergic transmission in the BLA on synaptic plasticity in the CA1 and the DG in response to theta stimulation. Muscimol microinfused into the BLA, either 10 min before or immediately after TS, impaired LTP in the vHC-CA1 pathway . Fisher post hoc test revealed significant differences between both CONT-MUS and CONT-MUS-post groups vs. CONT-VEH group (p < 0.01 for both) with no statically significant difference between CONT-MUS group vs. CONT-MUS-post group (p > 0.06), except for post-tetanic responses (1–5 min), which were significantly different (p < 0.05). Thus, muscimol microinfusion into the BLA tonically enhanced excitation in DG granular cells but not in CA1 pyramidal cells )see also In contrast, muscimol infused into the BLA, either 10 min before or immediately after TS, enhanced LTP in the PP-DG pathway , but enhanced LTP in the DG . ANOVA revealed a significant difference between groups subjected to BLA priming and control groups . When muscimol was infused after BLA stimulation, LTP was still impaired (last 10 min BLA-MUS: 254.47 ± 25.1%). ANOVA performed on these data revealed a significant effect of groups  = 3.49; p = 0.03) and groups–time interaction  = 2.98; p < 0.0001). Post hoc analyses showed a significant difference between CONT-VEH group and both BLA-MUS and BLA-VEH groups (p < 0.01) as well as between BLA-VEH and MUS-BLA groups (p < 0.01). No significant differences were observed between either CONT-VEH and MUS-BLA or BLA-VEH BLA-MUS groups . However, when muscimol was microinfused after BLA stimulation, LTP was significantly impaired, even compared to control . ANOVA performed on these data revealed a significant effect of groups  = 6.45; p = 0.002) and groups–time interaction  = 5.23; p < 0.0001). Post hoc analyses showed significant differences between the BLA-MUS group and the three other groups (p < 0.01) and no significant difference between BLA-VEH and MUS-BLA groups . As seen in BLA-BLA group exhibited stronger impairment in LTP in the CA1 as compared to the BLA-MUS group (p = 0.015) and the MUS-BLA group (p < 0.0001). In the DG, the BLA-BLA group was significantly different from both the BLA-MUS (p < 0.0001) and the MUS-BLA (p = 0.043) groups. However, post-tetanic potentiation (1–15 min post TS) was similar in the BLA-BLA group and the MUS-BLA groups (p > 0.05).We next examined whether priming can be performed more than once to affect metaplasticity  and whether this leads to accumulated priming effect, cancelation of the priming effect, or no change compared to the single priming, as well as how this compares to priming with muscimol. For this purpose, we added a group treated with two repeated BLA priming protocols delivered 10 min apart  housed in Plexiglas cages (five rats per cage) and maintained on a free-feeding regimen with a 12:12 h light/dark schedule. All experiments were performed during the light phase of the cycle at least one week after their arrival from the supplier . All experiments were carried out in accordance with the guidelines of the NIH and approved by the University of Haifa Animal Care and Use Committee. Rats were anesthetized with urethane  in saline (0.9 g/mL) and mounted onto a stereotaxic frame . The scalp was incised and retracted, and the head position was adjusted to place bregma and lambda in the same horizontal plane. Small burr holes (approximately 1.5 mm diameter) were drilled in the skull for the placement of electrodes and electrode/cannula assembly. During the course of the experiments, body temperature was monitored and maintained at 37 °C ± 0.5 by a regulated heating pad.A recording glass electrode  was stereotaxically positioned in the CA1 (4.2 mm posterior to bregma (AP), 2.5 mm from midline (ML), and ~2 mm dorsoventral (DV)) or in the DG . A bipolar concentric stimulating electrode  was inserted either in the controlateral ventral hippocampal commissure  for activating field potentials in the CA1 or in the ipsilateral perforant pathway  for activating field potentials in DG. The dorso-ventral location of the recording and stimulating electrodes was adjusted to maximize the amplitude of the evoked field potentials (EPs). A reference electrode consisting of a 100 μm coated wire was affixed to the skull in the area overlapping the nasal sinus.An electrode/cannula assembly was inserted in the ipsilateral BLA  for both drug microinfusion and stimulation. The electrode/cannula assembly was composed of a stimulating electrode made of two 45 µm diameter intertwined platinum-iridium wires  glued parallel to a 28-gauge stainless steel cannula with a separation of ~300 µm between tips. The assembly was inserted into the brain so that the electrode and the cannula were in a rostro-caudal alignment with the electrode placed rostral to the microinfusion cannula. CA1 and DG field potentials evoked by single pulses delivered to the vHC or the PP, respectively (0.1 msec rectangular monophasic pulses) were amplified (×1000) by a DAM50 Bio-Amplifier (World Precision Instruments (WPI), Sarasota, FL, US) displayed on an oscilloscope, digitized at 10 kHz (Cambridge Electronic Design Ltd (CED) Cambridge, UK), and saved on disk for off-line analysis . Baseline responses were established by means of a stimulation intensity (50–200 µA) sufficient to elicit a response representing ~25–30% of the maximal amplitude of the evoked field potentials. LTP was assessed by measuring the increase in the population spike amplitude (PS) of the EPs for both DG and CA1. In our previous study , we showA transmission, the GABAA receptor agonist muscimol  was used at a concentration of 0.05 µg/µL. Muscimol was dissolved in physiological saline, which was also used as control. An infusion volume of 0.3 µL  was delivered over a period of 3 min (at a rate of 0.1 µL/min) through a 28-gauge infusion cannula attached to a 10 µL Hamilton micro-syringe  via polyethylene (PE-20) tubing.To enhance GABACONT-VEH) or muscimol (CONT-MUS) was infused into the BLA, and a 10 min baseline recording session began after injection of the first 0.1 µL. The 10 min baseline recording was followed by application of theta-like high-frequency stimulation  to the vHC or the PP. After TS, responses to test pulse stimuli were recorded every 30 s for 1 h. In the third group (CONT-MUS post TS), muscimol was injected after TS, followed by a 1 h recording.To study the effect of manipulating BLA GABA transmission on LTP in the vHC-CA1 and the PP-DG pathways, three baseline and three priming groups of animals were used for each pathway . In the (BLA-VEH) or muscimol (MUS-BLA) recording was followed by BLA stimulation  30 s before TS to vHC or PP. Responses to low-frequency baseline pulses were then collected every 30 sec for 1h. In the third group (BLA-MUS), the BLA was stimulated prior to the 10 min muscimol infusion, followed by TS to the vHC or the PP and 1 h recording.In the priming groups, the 10 min baseline saline Muscimol was injected at a volume of 0.3 µL based on preliminary studies and the literature ,42,43 toAfter completion of the study, animals were transcardially perfused with physiological saline, followed by 10% buffered formalin. Brains were post-fixed in formalin-saccharose 30% solution for at least 3 days, frozen, and cut coronally on a sliding microtome into 50 µm sections. The sections were mounted on a gelatin-coated slide and stained with cresyl violet for microscopic examination of electrode/cannula assembly placement in the BLA.The amplitude of the PS of the EPs was expressed as the mean percentage (±SEM) of the individual basal values of animals for each group. Group differences were analyzed by ANOVA and Fisher post hoc tests (Statview).In the present study, we demonstrated that muscimol, a selective GABAA receptor (GABAAR) agonist, infused into the BLA differentially altered excitation and plasticity in the DG and the CA1. Priming of the BLA with muscimol enhanced excitation and plasticity in the DG but not in the CA1, thereby mimicking the previously reported effect of electrical priming of the BLA ,26. HoweThe results support the notion that the BLA modulates hippocampal activity and plasticity in a region-specific manner, with considerably different impact in the DG and the CA1 ,26. ThesWe recently demonstrated that inducing a highly selective lasting alteration of BLA activity by way of viral vectors aiming at selectively altering GABAergic synapses at the axon initial segment of principal BLA neurons affected both medial Prefrontal Cortex (mPFC)-dependent behaviors and amenability of the mPFC to induced plasticity . InteresThe present findings further emphasize the potential contribution of activity in GABAergic interneurons to the formation of a shift in the way the BLA modulates memory formation in other brain regions. Sources of GABAergic input to the BLA principal pyramidal-like glutamatergic projection neurons arise from at least two distinct populations of interneurons. The local-circuit neurons, which are scattered throughout the BLA, and the intercalated cell masses (ITCs) surrounding the BLA ,47,48. TIn our previous study, we showed that varying BLA stimulation patterns resulted in differential alterations in serum corticosterone level (CORT), with higher CORT levels being positively correlated with LTP magnitude in DG but not in CA1 . LikewisInhibition or inactivation of the BLA reduces anxiety and promotes hippocampal-dependent memory and memory processes . In contUnder resting conditions, the amygdala exhibits low neuronal firing due to this particularly high inhibitory tone exerted by the large GABAergic interneuronal network . Either While GABAARs are localized to the soma of pyramidal cells and interneurons, a study by Cunha and colleagues (2010)  suggestsIt is important to note that a general increased excitation, when combined with more specific and selective increased activity of some interneurons (due to BLA stimulation or to inhibition of inhibition by muscimol), can be expected to lead not to general enhanced effects of the BLA or suppression of its activity but rather to altered processing within the BLA, which modifies the modulation by the BLA of target areas, such as the hippocampus. However, BLA and muscimol may also activate different mechanisms, which may lead to similar consequences with respect to the impact on DG and CA1 excitation and LTP. The results obtained with our double priming protocol support such possibility. We showed that two sets of BLA stimulation (double priming) did not yield the same impact on CA1 and DG LTP as the BLA-muscimol protocol . This po


The current study explored changes in depression and anxiety symptoms from before the pandemic to soon after it first peaked in Spring 2020 in a sample of adolescents and young adults (N = 451) living in Long Island, New York, an early epicenter of COVID-19 in the U.S.The coronavirus [coronavirus disease 2019 (COVID-19)] pandemic has introduced extraordinary life changes and stress, particularly in adolescents and young adults. Initial reports suggest that depression and anxiety are elevated during COVID-19, but no prior study has explored changes at the Depression (Children's Depression Inventory) and anxiety symptoms (Screen for Child Anxiety Related Symptoms) were assessed between December 2014 and July 2019, and, along with COVID-19 experiences, symptoms were re-assessed between March 27th and May 15th, 2020.Across participants and independent of age, there were increased generalized anxiety and social anxiety symptoms. In females, there were also increased depression and panic/somatic symptoms. Multivariable linear regression indicated that greater COVID-19 school concerns were uniquely associated with increased depression symptoms. Greater COVID-19 home confinement concerns were uniquely associated with increased generalized anxiety symptoms, and decreased social anxiety symptoms, respectively.Adolescents and young adults at an early epicenter of the COVID-19 pandemic in the U.S. experienced increased depression and anxiety symptoms, particularly amongst females. School and home confinement concerns related to the pandemic were independently associated with changes in symptoms. Overall, this report suggests that the COVID-19 pandemic is having multifarious adverse effects on the mental health of youth. The novel coronavirus [coronavirus disease 2019 (COVID-19)] pandemic has disrupted the lives of individuals around the world. In the U.S., city and state-wide shelter-in-place orders were implemented across the country as early as March 2020, forcing most Americans to adjust to new circumstances (e.g. studying and working from home), take on new roles , and deal with new or exacerbated hardship . The nature and full extent of the impact of COVID-19 is only beginning to be understood as this unprecedented crisis unfolds.An emerging literature suggests that depression and anxiety symptoms may be elevated during the COVID-19 pandemic, and that certain populations  are more vulnerable to worsening mental health during the pandemic .The iPANDA project is a longitudinal investigation of developmental trajectories of neural reward sensitivity and psychopathology in a community sample of 317 girls from Long Island, New York  were recruited using a commercial mailing list of families with a 3-year-old child living in a 20-mile radius of Stony Brook University. Eligibility criteria included having a 3-year-old child with no significant medical disorder or developmental disability and a biological parent with the ability to read and write English. Following the age 3 assessment, families were invited to participate in follow-up assessments when their child was approximately age 6, 9, 12, 15, and 18. To increase the diversity of the sample, 50 children from underrepresented minority groups were added to the sample in the age 6 wave. At the time of the COVID-19 pandemic, the SBTS had just begun its sixth wave (age 18) of data collection. Therefore, the pre-COVID-19 assessment was identified as the fifth wave of data collection .The SBTS is a longitudinal study designed to explore early precursors to depression and anxiety disorders in a community sample of youth from Long Island, NY , sometimes (1), and often (2). There are 5 subscales that measure different anxiety symptoms that parallel anxiety disorders classified in the DSM, including panic/somatic symptoms, generalized anxiety, separation anxiety, social anxiety, and school phobia. The SCARED has demonstrated adequate psychometric properties in both clinical and non-clinical samples . The infection concerns composite included two items probing concern about and perceived likelihood of becoming infected with COVID-19, rated on a Likert-scale from 0  to 4 (extremely). Scores ranged from 0 to 8 . The school concerns composite included a checklist of four school-related concerns . Scores ranged from 0 to 4 . The home confinement concerns composite included a checklist of three concerns related to being largely restricted to home (e.g. experiencing cabin fever). Scores ranged from 0 to 3 . Finally, the basic needs concerns subscale included a checklist of three concerns related to having one's basic needs met (e.g. not having enough food or supplies). Scores ranged from 0 to 3 .The life changes composite included a checklist of 14 changes in life circumstances related to the pandemic . Scores ranged from 0 to 8  = 0.47, p < 0.001, were included as covariates in all analyses. Controlling for age at both assessments allowed us to account for not only age effects but also differences in the interval between assessments. Change in CDI and SCARED symptoms was examined via repeated measures analysis of covariance (ANCOVA) with time (pre-COVID-19 v. COVID-19) as a within-subject factor and age at the pre-COVID-19 and COVID-19 assessments as covariates. Repeated measures ANCOVA allowed us to test changes in symptoms as a function of time, while controlling for continuous covariates (age). Gender difference in symptom change was examined via mixed-design ANCOVA with time (pre-COVID-19 v. COVID-19) as a within-subject factor and gender [females (1) v. males (0)] as a between-subjects factor, and age at the pre-COVID-19 and COVID-19 assessments as covariates. Cohen's d, an effect size index of the magnitude of difference between means, is reported for symptom changes pre-COVID-19 to COVID-19 .Adolescence is a critical period for the emergence and exacerbation of depression and anxiety symptoms  v. males (0)], all other CDI and SCARED symptom residuals, and all pandemic experiences composites were included as independent variables, and the CDI or SCARED symptom residual of interest served as the dependent variable. Separate analyses were conducted for each CDI and SCARED residual as the dependent measure. All analyses were conducted in IBM SPSS Statistics 26.0.Associations between the pandemic experiences composites and change in CDI and SCARED symptoms was examined with a three-step analytical approach. First, CDI and SCARED symptom residuals were created by regressing COVID-19 symptoms on pre-COVID-19 symptoms. Residuals were used instead of raw change scores, as the latter is liable to problems with reliability and regression to the mean. Next, partial correlations were computed between the pandemic experiences composites and COVID-19 symptom residuals, adjusting for age at the pre-COVID-19 and COVID-19 assessments. Finally, multivariable linear regression was conducted to examine 2(101) = 102.03, p = 0.43 , including 295 (65.4%) females and 374 (82.9%) individuals who identified as White/non-Hispanic. Participant educational background included 305 (67.6%) who were currently in high school and 141 (31.3%) who were currently in college. Parental educational background included 289 (64.1%) participants with a parent who was a college graduate.Among the 713 participants who completed the pre-COVID assessment, 505 (70.8%) participated in the COVID-19 assessment, and 54 were missing symptom or pandemic experience data. Little's MCAR test suggested that listwise deletion of participants with incomplete COVID-19 assessment data was appropriate, χ Little, . The finn = 451), relative to those who did not complete the COVID-19 assessment or were excluded from analyses (n = 262), were more likely to be female, White/non-Hispanic, 74.8% v. 64.4%, χ2(1) = 6.81, p = 0.01, and have a parent who was college educated, 53.8% v. 49.2%, χ2(1) = 7.63, p = 0.01. Groups did not differ in age at the pre-COVID-19 assessment, t(711) = −0.48, p = 0.63.Attrition analyses indicated that participants included in the final analyses  = 6.22, p = 0.013 and panic/somatic symptoms, F = 4.80, p = 0.029, but not generalized or social anxiety symptoms. Post hoc analyses reveal that depression and panic/somatic symptoms increased for females only.We conducted sensitivity analyses controlling for study and all demographic variables. None of the results changed substantively when accounting for these variables.Prior studies using nationally representative samples have suggested that incidence of depression and anxiety increased following the COVID-19 pandemic in the U.S., which has been particularly impacted (Ettman et al., Findings of the current study suggest that the COVID-19 pandemic contributed to increased symptoms of generalized and social anxiety in youth living in Long Island, New York. Moreover, females also experienced increased depression and panic/somatic symptoms. Depression and anxiety typically increase from late childhood through early adulthood (Gruber et al., Consistent with research on prior outbreaks (Wenham et al., decreased social anxiety symptoms, indicating that, for some youth, the pandemic has provided a respite from social pressures. Further work is needed to determine how this will impact social development and anxiety after the pandemic.Across domains, pandemic experiences were associated with increases in psychiatric symptoms. Specifically, greater concern about contracting COVID-19 and school-related problems were associated with increases in both depression and all three types of anxiety symptoms, while experiencing more life changes and greater concern about being confined at home and meeting basic needs were associated with increased depression, panic/somatic symptoms and generalized anxiety. Multivariable analyses suggest that school concerns, including concerns about passing classes, juggling schoolwork with other responsibilities and online classes being poor quality, were uniquely associated with increases in depression symptoms. This is consistent with a study on adolescents in Australia during COVID-19 (Magson et al., This study possesses many strengths, including a longitudinal design capturing within-person changes in symptoms, assessment of a range of pandemic experiences, and a sample residing in one of the largest epicenters of the pandemic at the time of assessment. However, there were several limitations. The attrition rate raises concerns about generalizability of these findings. Participant recruitment was non-random, although the demographic make-up of the baseline sample is similar to census data for Suffolk County, NY (U.S. Census Bureau,